Molecular characteristics of common sandflies species (Diptera: Psychodidae) in Hainan province

doi:10.11853/j.issn.1003.8280.2018.01.004

Abstract

Abstract: Objective To identify the common species of sandflies in Haikou city based on morphological characteristics, and determine the species specific molecular identity. Methods Sandflies were collected overnight by light-trap monthly in the pigsty, chicken or duck coops or human dwellings around village and urban-rural fringe areas in Haikou city during August 2015 to July 2016, as well as in Sansha city in August, 2017. After counting, partial sandflies were randomly selected and classified by morphological characteristics. Then the genomic DNA was extracted from identified specimens, the mtDNA Cytb gene fragment was sequenced and analyzed to determine the species-specific molecular identity. The sandflies collected from Sansha city were identified by molecular marker, and the taxonomic status was discussed. Results A total of 377 sandflies samples were collected in Haikou city, which belonged to 4 species in 2 genera, as Sergentomyia bailyi Sinton, S.iyengari Sinton, S. squamipleuris Newstead, and Phleboto musstantoni Newstead. The sandflies adults were active throughout the year with the peak of density in June and November. The intraspecific genetic distance of mtDNA Cytb gene sequences among the 4 species was from 0 to 0.046. The range of the interspecific distance among the 9 species (including 5 species mtDNA Cytb gene sequences downloaded from NCBI database) was from 0.112 (P. wui with P. chinensis) to 0.219 (P. stantoni and P. caucasicus). All sandflies specimen collected from Sansha city were S. bailyi by molecular identity. The most genetic distance was 0.049 between Haikou and Sansha population. Conclusion Sandflies were active throughout the year in Haikou city, including two peaks of density. The identity of mtDNA Cytb gene sequences of 4 common sandflies in Hainan province was identified. There was obvious genetic divergence in S. bailyi populations.

Key words: Sandfly, Classification, Molecular identity, Mitochondrial DNA, Hainan province

摘要： 目的依据形态特征对海南省常见白蛉种类进行分类，确定该地常见白蛉的种特异分子特征。方法利用诱虫灯法，于2015年8月至2016年7月每月在海南省海口市农村和城乡结合部的猪圈、鸡鸭圈或人房等环境，2017年8月在三沙市宿舍周围的院落、废弃的猪圈等环境通宵采集白蛉，统计白蛉数量。随机挑取白蛉样本依据形态学特征进行分类鉴别，对应标本提取基因组DNA，测定和分析线粒体DNA（mtDNA）细胞色素b（Cytb）基因片段序列，确定种特异的分子特征。对采自海南省三沙市的白蛉应用种特异分子特征鉴定种类，并探讨其分类地位。结果在海口市共捕获白蛉377只，隶属于2属4种，即贝氏司蛉、应氏司蛉、鳞胸司蛉和施氏白蛉；白蛉全年均有活动，密度高峰出现在6、11月。4种白蛉的mtDNA Cytb基因序列在个体间的遗传距离为0~0.046，在我国分布的9种（5种为网络下载）白蛉间的差异范围为0.112~0.219。应用mtDNA Cytb基因序列分析显示，在三沙市采集的白蛉均为贝氏司蛉，与海口市贝氏司蛉间的遗传差异最大为0.049。结论白蛉在海口市全年均有活动，有2个密度高峰；确定了海南省常见4种白蛉的mtDNA Cytb基因分子特征；基于分子特征的贝氏司蛉已出现群体分化。

关键词: 白蛉, 分类, 分子特征, 线粒体DNA, 海南省

CLC Number:

R384.9

CHEN Hui-ying, LIN Yi, CHEN Han-ming, LIU Yi, LI Kai-li, GAO Jing-peng, CHEN Xue-wen, CAI Fang, LIU Chao, YANG Xin-yan, YANG Zhen-zhou, PENG Heng, MA Ya-jun. Molecular characteristics of common sandflies species (Diptera: Psychodidae) in Hainan province[J]. Chines Journal of Vector Biology and Control, 2018, 29(1): 15-19.

陈辉莹, 林怡, 陈翰明, 刘义, 李凯利, 高景鹏, 陈学文, 蔡芳, 刘超, 杨新艳, 杨振洲, 彭恒, 马雅军. 海南省常见白蛉种特异分子特征的研究[J]. 中国媒介生物学及控制杂志, 2018, 29(1): 15-19.

References

[1] Depaquit J. Molecular systematics applied to Phlebotomine sandflies:review and perspectives[J]. Infect Genet Evol, 2014,28:744-756.
[2] Service MW. Medical entomology for students[M]. 3rd ed. Liverpool:Cambridge University Press,2004:90-98.
[3] 熊光华,金长发,管立人. 中国的白蛉[M]. 北京:科学出版社,2016:15-25.
[4] 杨曼尼,马雅军. 我国白蛉的分类现状[J]. 国际医学寄生虫病杂志,2008,35(1):46-49.
[5] Zhang LM, Leng YJ. Eighty-year research of Phlebotomine sandflies (Diptera:Psychodidae) in China (1915-1995). Ⅱ. Phlebotomine vectors of Leishmaniasis in China[J]. Parasite, 1997,4(4):299-306.
[6] 管立人. 中国白蛉(双翅目:毛蛉科)调查研究工作的展望[J]. 中国寄生虫学与寄生虫病杂志,2013,31(4):310-314.
[7] 冷延家. 在海南岛继续发现的蛉种与新种白蛉:方亮白蛉Phlebotomus fanglianensis sp. nov.的记述[J]. 昆虫学报, 1964,13(1):118-128.
[8] 杨明磊,李逸明,陈辉莹,等. 我国西沙群岛吸血昆虫的种群动态和鼠形动物病原感染检测[J]. 第二军医大学学报, 2016,37(3):355-359.
[9] Kumar NP, Srinivasan R, Jambulingam P. DNA barcoding for identification of sand flies (Diptera:Psychodidae) in India[J]. Mol Ecol Resour,2012,12(3):414-420.
[10] de Souza Pinto I, das Chagas BD, Rodrigues AAF, et al. DNA barcoding of neotropical sand flies (Diptera, Psychodidae, Phlebotominae):species identification and discovery within Brazil[J]. PLoS One,2015,10(10):e0140636.
[11] Polseela R, Jaturas N, Thanwisai A, et al. Towards monitoring the sandflies (Diptera:Psychodidae) of Thailand:DNA barcoding the sandflies of Wihan Cave, Uttaradit[J]. Mitochondrial DNA A DNA Mapp Seq Anal,2016,27(5):3795-3801.
[12] Kocher A, Gantier JC, Gaborit P, et al. Vector soup:high-throughput identification of Neotropical phlebotomine sand flies using metabarcoding[J]. Mol Ecol Resour,2017,17(2):172-182.
[13] Dokianakis E, Tsirigotakis N, Christodoulou V, et al. DNA sequencing confirms PCR-RFLP identification of wild caught Larroussius sand flies from crete and cyprus[J]. Acta Trop, 2016,164:314-320.
[14] Gajapathy K, Tharmasegaram T, Eswaramohan T, et al. DNA barcoding of Sri Lankan phlebotomine sand flies using cytochrome c oxidase subunitⅠ reveals the presence of cryptic species[J]. Acta Trop,2016,161:1-7.
[15] Esseghir S, Ready PD, Ben-Ismail R. Speciation of Phlebotomus sandflies of the subgenus Larroussius coincided with the late Miocene-Pliocene aridification of the Mediterranean subregion[J]. Biol J Linn Soc,2000,70(2):189-219.
[16] 冷廷家. 贝氏白蛉及其亚种分类的研究[J]. 暨南大学学报:自然科学版,1980,1(2):23-30.
[17] 高景鹏,李凯利,陈辉莹,等. 菊酯类杀虫剂对海口市白纹伊蚊的效果测定[J]. 中国热带医学,2017,17(9):867-870.
[18] Zhang L, Ma YJ, Xu JN. Genetic differentiation between sandfly populations of Phlebotomus chinensis and Phlebotomus sichuanensis (Diptera:Psychodidae) in China inferred by microsatellites[J]. Parasit Vectors,2013,6(1):115.
[19] Zhang L, Ma YJ. Identification of Phlebotomus chinensis (Diptera:Psychodidae) inferred by morphological characters and molecular markers[J]. Entomotaxonomia,2012,34(1):71-80.
[20] Parvizi P, Naddaf SR, Alaeenovin E. Molecular typing and phylogenetic analysis of some species belonging to Phlebotomus (Larroussius) and Phlebotomus (Adlerius) subgenera (Diptera:psychodidae) from two locations in Iran[J]. Iran J Arthropod Borne Dis,2010,4(2):1-10.
[21] Krüger A, Strüven L, Post RJ, et al. The sandflies (Diptera:Psychodidae, Phlebotominae) in military camps in northern Afghanistan (2007-2009), as identified by morphology and DNA "barcoding"[J]. Ann Trop Med Parasitol,2011,105(2):163-176.
[22] Berdjane-Brouk Z, Koné AK, Djimdé AA, et al. First detection of Leishmania major DNA in Sergentomyia (Spelaeomyia) darlingi from cutaneous leishmaniasis foci in Mali[J]. PLoS One,2012,7(1):e28266.
[23] Gajapathy K, Peiris LB, Goodacre SL, et al. Molecular identification of potential leishmaniasis vector species within the Phlebotomus (Euphlebotomus) argentipes species complex in Sri Lanka[J]. Parasit Vectors,2013,6(1):302.
[24] Kasap OE,Vot'pka J,Alten B. The distribution of the Phlebotomus major complex (Diptera:Psychodidae) in Turkey[J]. Acta Trop,2013,127(3):204-211.
[25] Bounamous A, Lehrter V, Hadj-Henni L, et al. Limits of a rapid identification of common Mediterranean sandflies using polymerase chain reaction-restriction fragment length polymorphism[J]. Mem Inst Oswaldo Cruz,2014,109(4):466-472.
[26] Depaquit J,Randrianambinintsoa FJ,Jaouadi K,et al. Molecular and morphological systematics of the sandfly Sergentomyia (Sintonius) clydei Sinton, 1928 and questions about its record in the Seychelles[J]. Infect Genet Evol,2014,21:41-53.
[27] Ebrahimi S,Bordbar A, Parvizi P. Genetic dynamics in the sand fly (Diptera:Psychodidae) nuclear and mitochondrial genotypes:evidence for vector adaptation at the border of Iran with Iraq[J]. Parasit Vectors,2016,9(1):319.
[28] 张丽,马雅军. 基于线粒体DNA的我国中华白蛉群体遗传分化研究[J]. 中国热带医学,2016,16(10):947-952.
[29] Yogeswari S,Srinivasan R. A note on variations in morphological features of the Phlebotomine sand fly Sergentomyia bailyi (Diptera:Psychodidae) in a population from Pondicherry UT, India[J]. J Med Entomol,2016,53(3):712-716.
[30] Tharmatha T, Gajapathy K, Ramasamy R, et al. Morphological and molecular identification of cryptic species in the Sergentomyia bailyi (Sinton,1931) complex in Sri Lanka[J]. Bull Entomol Res,2017,107(1):58-65.